Peptides
Please refer to the current literature for information on the cleaning and use of peptides.
A concentration of at least 5–10 µg/ml should be used for injection, because some peptides rapidly degrade in the cells after injection.
Oligonucleotides
Purification The purification of oligonucleotide solutions is very important. Cleaning with gel or HPLC is recommended.
Like DNA, oligonucleotides are dissolved in bidist. water after purification.
A concentration of 1–2 µg/ml should be used for injection of antisense oligonucleotides with 10–20 bases.
Note: Injected oligos accumulate easily in the nucleus.
Literature
[1] Pepperkok, R., Schneider, C., Philipson, L., and Ansorge, W. (1988). Single cell assay with an automated capillary microinjection system. Exp.Cell Res. 178, 369-376.
[2] Pepperkok, R., Scheel, J., Horstmann, H.,Hauri, H.P., Griffiths, G., and Kreis,T. E. (1993b). ß-COP is essential for biosynthetic membrane transport from the endplasmic reticulium to the Golgi complex in vivo. Cell 74, 71-82.
[3] Pepperkok, R., Saffrich, R., and Ansorge, W. (1994). Computer-Automated Capillary Microinjection of Macromolecules into Living Cells. Cell Biology: A Laboratory Handbook, CSH Laboratory Press
[4] Antibodies: A laboratory manual (1988). Harlow, E. and Lane, D. ed., CSH Laboratory Press.
[5] Proctor, G.N. (1992). Microinjection of DNA into mammalian cell in culture: Theory and practice. Methods Mol. Cell. Biol. 3, 209-231.
[6] Sambrook, Fritsch, Maniatis (1989). Molecular Cloning: A laboratory manual. CSH Laboratory Press.
We thank Dr. Rainer Pepperkok, Université de Genéve, Département de Biologie Cellulaire Sciences III, CH 1211 Genève 4, for providing us with data.
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